RT-PCR for full-length IL-17RC and TaqMan probe detection.
Total RNA was extracted with the RNeasy Mini kit (QIAGEN) and reversed-transcribed to generate cDNA with the High-Capacity cDNA Reverse Transcription kit (Invitrogen). Platinum High-Fidelity Taq DNA Polymerase (Invitrogen) was then used to amplify the full-length IL17RC cDNA.
The following primers were used: F, 5′-CTGAAGAGGGATTCCAGCCC-3′ and R, 5′-GTAGAAAAACAGCGTCTGCC-3′. TaqMan probes for IL-17RC (Invitrogen) were used to detect mRNA, and the results were normalized against those for GUS (Human GUSB Endogenous Control VIC/MGB Probe; Primer Limited, Invitrogen).